Extended Data Figure 1: AcrIIA2 and AcrIIA4 specifically interact with sgRNA-bound SpyCas9.
From: Structural basis of CRISPR–SpyCas9 inhibition by an anti-CRISPR protein
Binding assays were carried out between the anti-CRISPR protein of AcrIIA2 or AcrIIA4 and the GST-tagged CRISPR protein of SpyCas9 or NmeCas9 in the presence or absence of cognate sgRNA. The purified GST–SpyCas9 or GST–NmeCas9 protein was first bound to glutathione sepharose beads in the presence or absence of sgRNA, and then the beads were incubated them with AcrIIA2 or AcrIIA4 protein. After extensive washing, the bound proteins were visualized by Coomassie staining following SDS–PAGE. Data shown are the representative of three replicates.
